LOCUS       sequence-373911-        2709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP from PR promoter in E.coli Dh5alpha cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2709)
  AUTHORS   Mukhopadhyay S, Bagh S
  TITLE     A microgravity responsive synthetic genetic device in Escherichia 
            coli.
  JOURNAL   Biosens Bioelectron. 2020 Nov 1;167:112462. doi: 
            10.1016/j.bios.2020.112462. Epub 2020 Aug 1.
  PUBMED    32781386
REFERENCE   2  (bases 1 to 2709)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biosens 
            Bioelectron."; date: "2020-11-1"; volume: "167:112462. doi"; pages: 
            " 10.1016/j.bios.2020.112462. Epub 2020 Aug 1"
COMMENT     Sequence Label: P_EGFP_AC
FEATURES             Location/Qualifiers
     source          1..2709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             77..88
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(95..97,98..100,101..814)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(95..97,98..100,101..811)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(140..161)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(401..420)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     748..769
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      826..912
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(947..964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1081..1626)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1161..1180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1740..1834)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1855..2514)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2429..2448
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2515..2617)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2683..2701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcacctcg agtaacaccg tgcgtgttga ctattttacc tctggcggtg ataatggttg
       61 catgtacgaa ttcattaaag aggagaaagg taccatggtg agcaagggcg aggagctgtt
      121 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      181 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      241 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      301 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      361 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      421 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      481 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      541 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
      601 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
      661 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
      721 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
      781 gatcactctc ggcatggacg agctgtacaa gtaatctaga ggcatcaaat aaaacgaaag
      841 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      901 agtaggacaa atccgccgcc ctagacctag ggatatattc cgcttcctcg ctcactgact
      961 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
     1021 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
     1081 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
     1141 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     1201 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     1261 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     1321 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1381 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     1441 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     1501 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     1561 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     1621 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     1681 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgttact agtgcttgga
     1741 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     1801 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
     1861 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
     1921 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     1981 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     2041 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     2101 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     2161 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     2221 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     2281 tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     2341 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     2401 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     2461 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     2521 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     2581 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccagatatc
     2641 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     2701 ccctttcgt
//
